Scientia Agricultura Sinica ›› 2013, Vol. 46 ›› Issue (9): 1903-1914.doi: 10.3864/j.issn.0578-1752.2013.09.018

• ANIMAL SCIENCE·RESOURCE INSECT • Previous Articles     Next Articles

Optimization of Incubation of Sperm and Lentivirus and Transgenic Pigs Production

 CAI  Wei-Guang, XI  Qian-Yun, XIAO  Min, GUAN  Li-Zeng, MENG  Fan-Ming, SHU  Gang, ZHU  Xiao-Tong, JIANG  Qing-Yan, WU  Zhen-Fang, ZHANG  Yong-Liang   

  1. College of Animal Science, South China Agricultural University/Guangdong Provincial Key Lab of Agoanimal Genomics and Molecular Breeding, Guangzhou 510642
  • Received:2012-05-06 Online:2013-05-01 Published:2012-09-05

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Abstract: 【Objective】A new method was developed for producing transgenic pig using incubation of lentivirus and sperm. 【Method】Effects of sperm number, amount of lentivirus, incubation time and incubation temperature on sperm activity were assessed by the orthogonality design. PCR and southern blotting were used to analyze the transgene.【Result】The optimized incubation was incubation of 100 µL lentivirus (5×105cfu) with 1.0×107 sperms per mL for 30 min at 17℃. Using this procedure sperms were artificially inseminated into sows. PCR was used to detect the offspring. Fourteen of 49 (14/49) piglets were PCR-positive. Four of 8 PCR-positive pitlets showed foreign gene intergration revealed by using southern blotting.【Conclusion】Incubation indexes of lentiviruse and sperm optimized in this study are that 100 µL lentivirus (5×105cfu) with 1.0×107 sperms per mL for 30 min at 17℃, and transgenic pigs were produced by using this procedure. Foreign gene intergration were revealed.

Key words: lentivirus , SMGT , transgenic pig

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